Purification of a Modified Flagellar Export Apparatus

改良鞭毛出口装置的纯化

• 批准号: 8394244
• 负责人: JONATHAN L MCMURRY
• 金额: $ 2.29万
• 依托单位: KENNESAW STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-01 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8394244
• 关键词: Address; Affect; Affinity; Award; Bacterial Infections; Binding; Biochemical; Biological Assay; Biological Models; Biological Process; Biomedical Research; Biosensing Techniques; Chemotaxis; Collaborations; Complement; Complex; Detergents; Development; Dissociation; Dysentery; Environment; Event; Filament; Flagella; Funding; Genes; Genetic; Goals; In Vitro; Institution; Interferometry; Kinetics; Liposomes; Measures; Membrane; Mentors; Methods; Molecular; Molecular Biology; Optics; Pathogenesis; Positioning Attribute; Preparation; Process; Progress Reports; Protein Export Pathway; Proteins; Protons; Publications; Recruitment Activity; Research; Research Personnel; Research Training; Salmonella enterica; Signal Transduction; Structure; Students; Surveys; System; Techniques; Training; Typhoid Fever; United States National Institutes of Health; Variant; Vesicle; Virulence; Work; assay development; career; cell motility; improved; in vivo; instrumentation; kinetosome; meetings; member; nanomachine; pathogenic bacteria; programs; protein protein interaction; reconstitution; retinal rods; stoichiometry; success; tool; undergraduate research

DESCRIPTION (provided by applicant): The bacterial flagellum is a self-assembling, proton-powered rotary nanomachine that drives motility, allowing chemotaxis. It contains a dedicated type III secretion (T3S) apparatus. Homologous T3S systems are used by pathogenic bacteria to effect virulence, underlying many bacterial diseases such as typhoid fever and bacterial dysentery. Using Salmonella enterica as a model system, we will investigate dynamic interactions among flagellar proteins that allow membrane translocation of export substrates through use of molecular biology, genetics and biochemical techniques including optical biosensing. The specific aims are to complete a kinetic survey of interactions among export apparatus proteins using optical biosensing as the primary tool, to examine higher-than-pairwise interactions that better reflect in vivo dynamics and to further the development of an in vitro T3S assay. Eventual development of an in vitro export assay will be of tremendous significance because it would allow us to examine hypotheses addressing T3SS function currently out of reach. Since the flagellar export apparatus is highly similar to virulence T3S systems, the project will contribute significantly to our understanding of bacterial pathogenesis. AREA program goals of enhancing the research environment and exposing students to meritorious research will be well-served by this project.

描述(申请人提供)：细菌鞭毛是一种自组装的、质子动力的旋转纳米机器，它驱动运动，允许趋化性。它包含一个专用的III型分泌物(T3s)装置。病原菌使用同源的T3s系统来产生毒力，导致许多细菌性疾病，如伤寒和细菌性痢疾。以沙门氏菌为模型系统，我们将利用分子生物学、遗传学和包括光学生物传感在内的生化技术来研究鞭毛蛋白之间的动态相互作用，这些鞭毛蛋白允许出口底物的膜转运。其具体目的是以光学生物传感为主要工具完成出口器官蛋白质之间相互作用的动力学调查，检查更好地反映体内动态的高于成对的相互作用，并进一步发展体外T3S分析。体外出口试验的最终发展将具有巨大的意义，因为它将使我们能够检验目前尚不能实现的T3SS功能的假说。由于鞭毛输出装置与毒力T3S系统高度相似，该项目将对我们理解细菌的发病机制做出重大贡献。这个项目将很好地服务于改善研究环境和让学生接触到有价值的研究的区域计划目标。